Inappropriate epigenetic modifications of gene expression are associated with malignant phenotype and tumor progression. Regulation of gene expression is mediated by several mechanisms such as DNA methylation, ATP-dependent chromatin remodeling, and post-translational modifications of histones. Enzymes responsible for the reversible acetylation/deacetylation processes are histone acetyltransferases (HATs) and histone deacetylases (HDACs), respectively. Inhibitors of histone deacetylase (HDAC) enzymes have gained prominence as an emerging class of anticancer agents. We synthesized a series of hydroxamic acid-based HDAC inhibitors, characterized by a biphenyl-4-yl-acrylohydroxamic acid skeleton. The compounds were designed based on the hypothesis that the phenyl ring, extended from the hydroxamic acid via a double bond, could be suitable to occupy the narrow tube-like pocket of the HDAC active site. On the contrary, the cap structure appeared to accommodate in a large cavity, without a significant steric clash. Thus, a modification of the cap group appeared the most promising strategy to optimize drug-target interaction. Molecular docking was used to predict the optimal conformation of different molecules in the active site of a representative isoform from class I (HDAC2, sharing 85% sequence identity and 93% of sequence homology with HDAC1) and class II (HDAC6). From the results of docking studies three potential candidates with increased drug-target interactions were selected. All of them showed an activity towards HDAC1, 2 and 6 in the low µM range, estimated through a fluorimetric activity assay [2]. One of the compounds was further tested in vitro and in vivo in a colon carcinoma model and showed significant proapoptotic and antitumor activities.